DNA insert from 4114 vector PHP27118
The DNA insert contains four gene cassettes: Bacillus thuringiensis cryF, B. thuringiensis cry34Ab1, B. thuringiensis cry35Ab1 and Streptomyces viridochromogenes phosphinothricin N-acetyltransferase (pat). Transcription of the cryF is under control of a Zea mays ubiquitin promoter and an Agrobacterium tumefaciens ORF25 terminator. A Z. mays ubiquitin intron 1 sequence is also present to enhance the expression of cry1F. Transcription of the cry34Ab1 is under control of a Z. mays ubiquitin promoter and Solanum tuberosum proteinase inhibitor II terminator. A Z. mays ubiquitin intron 1 sequence is also present to enhance the expression of cry34Ab1. Transcription of cry35Ab1 is under control of Triticum aestivum peroxidase promoter and S. tuberosum proteinase inhibitor II terminator. Transcription of pat is under control of Cauliflower mosaic virus (CaMV) 35S promoter and CaMV 35S terminator.

Note:
- The ubiquitin gene promoter element is made up of  the promoter region (900bp) and the 5' untranslated region (83bp) of the ubiquitin gene.
- Transcription is expected to be occur at elevated levels for the present gene cassettes.
- Southern blot analyses indicated that a single, intact transformation cassette was inserted into the genome of 4114 maize with no integration of the backbone of plasmid PHP27118.


DNA insert from MON89034 vector PV-ZMIR245
Two insecticidal protein expression cassettes were inserted into the genome. Bacillus thuringiensis cry1A.105 expression is under the control of the CaMV 35S enhanced promoter, which first transcribes wheat (Triticum aestivum) 5' untranslated region of the chlorophyll a/b-binding protein (cab) and a rice actin 1 intron before transcribing cry1A.105. Transcription terminates at the wheat heat shock protein 17.3 terminator. Expression of the B. thuringiensis cry2Ab2 starts at the Figwort mosaic virus 34S promoter, which transcribes the Zea mays heat shock protein 70 (hsp70), then the Z. mays transit peptide and the cry2Ab2 coding sequence, before terminating at the nos terminator.

Note:
- The Cry2Ab2 coding sequence was modified for optimal expression in plants.
- South blot analysis confirmed that single insertions of both cry2Ab2 and cry1A.105, as well as no vector backbone were present and in the parent.
- A deletion removed the duplicated enhancer elements compared to the original CaMV 35S enhanced promoter in PV-ZMIR245.
- The selectable marker, nptII, cassette was bred out of the parental line and became not associated with this transformation event.


DNA insert from MON87411 vector PV-ZMIR10871
The MON87411 genome contains three cassettes: an RNA interference (RNAi) cassette targeting Diabrotica virgifera virgifera, Bacillus thuringiensis cry3Bb1 and Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (cp4-epsps).

Transcription of the RNAi cassette commences from the Cauliflower mosaic virus 35S enhanced promoter and terminates at the Pisum sativum ribulose bisphosphate carboxylase small chain 2 terminator. The transcript initially contains a Zea mays heat shock protein 70 intron, which contributes to enhanced expression in vegetative tissues of the plant, and two partial coding sequences of the D. virgifera virgifera Snf7p gene, which encodes the SNF7 subunit of the ESCRT-III complex. The two Snf7p sequences are in an inverted orientation, separated by a 150-nucleotide intervening sequence, which allows base pairing between the inverted sequences and hairpin RNA formation post-transcription, which then triggers an RNAi response. Due to RNAi processing, small interfering RNA molecules (roughly 21-23 nucleotides in length) will be produced and thus no translation into protein will occur from this cassette.

Transcription of the cry3Bb1 is under control of the Z. mays physical impedance induced protein promoter and Triticum aestivum (wheat) heat shock protein 17.3 terminator. The transcript also contains a wheat 5' untranslated leader from chlorophyll a/b-binding protein and Oryza sativa actin 1 intron for enhanced expression of the transgene. Expression of cp4-epsps is under control of an O. sativa alpha tubulin promoter and terminator. The transcript additionally contains Arabidopsis thaliana chloroplast targeting peptide 2 to sequester the protein to the chloroplast.

Note:
- Sequencing, PCR and bioinformatic analyses indicate that a single, intact insertions of the three gene cassettes occurred in the parental line.
- No plasmid backbone was detected.


DNA insert from DAS40278 vector pDAS1740
The LMO was generated using the Whiskers-mediated transformation method. Sphingobium herbicidovorans aryloxyalkanoate dioxygenase-1 (aad-1) is under the control of Zea mays ubiquitin gene promoter and Z. mays root preferential cationic peroxidase terminator. Elevated levels of transcription are expected to occur due to the constitutive nature of the ubiquitin promoter.

Note:
- The aad-1 coding sequence was optimized for expression in the plant.
- Southern blot analysis indicated that a single complete copy of the transformation cassette was stably integrated into the host genome at a single locus
- No integration of the vector backbone occurred.


Kindly refer to the parental LMO records for more information.